LOCUS       sequence-349071-        6136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Develop a family of genetically encoded fluorescent biosensors for 
            nicotinic agonists, termed iDrugSnFRs, thus enabling optical 
            subcellular pharmacokinetics for nicotinic agonists.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6136)
  AUTHORS   Nichols AL, Blumenfeld Z, Fan C, Luebbert L, Blom AE, Cohen BN, 
            Marvin JS, Borden PM, Kim CH, Muthusamy AK, Shivange AV, Knox HJ, 
            Campello HR, Wang JH, Dougherty DA, Looger LL, Gallagher T, Rees DC,
            Lester HA
  TITLE     Fluorescence activation mechanism and imaging of drug permeation 
            with new sensors for smoking-cessation ligands.
  JOURNAL   eLife. 2022 Jan 4;11. pii: 74648. doi: 10.7554/eLife.74648.
  PUBMED    34982029
REFERENCE   2  (bases 1 to 6136)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "eLife."; 
            date: "2022-01-4"; volume: "11. pii: 74648. doi"; pages: " 
            10.7554/eLife.74648"
COMMENT     Sequence Label: pAAV9-hSyn-iDianiSnFR_PM
FEATURES             Location/Qualifiers
     source          1..6136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     sig_peptide     27..86
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     539..560
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1665..1694
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1915..2503
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1968..1988)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2386..2397)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2506..2522)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2507..2523)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2552..2673
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2563..2582)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2617..2636
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2835..2964
                     /label=AAV2 ITR
     rep_origin      complement(3201..3789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3281..3300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3960..4751,4752..4820))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4583..4602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4821..4925)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4951..5406)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5088..5109)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5300..5319
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5478..5607
                     /label=AAV2 ITR
     promoter        5689..6136
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 tcgaattcgg cttggggata tccaccatgg agacagacac actcctgcta tgggtactgc
       61 tgctctgggt tccaggttcc actggtgaca gatctgcgaa cgacaccgta gttgtgggct
      121 cgatcgattt tacagaaggg attatcgtcg caaacatggt ggcagagatg attgaggcgc
      181 atacagacct taaggtggtt cgcaaactga accttggcgg gcgtaacgtt aactttgaag
      241 ccattaaacg cggaggtgcg aataatggta ttgacattta cgtggagtac actgggtccg
      301 gtcttgtgga tattctgggg ttcccggagc cgaacgtcta tatcaccgcc gacaagcaga
      361 agaacggcat caaggcgaac ttcaagatcc gccacaacat ggaggacggc agcgtgcagc
      421 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
      481 accactacct gagcacccag tccgtgctga gcaaagaccc caacgagaag cgcgatcaca
      541 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
      601 agggcggtac cggagggagc atgagcaagg gcgaggagct gttcaccggg gtggtgccca
      661 tcctggtcga gctggacggc ggcgtaaacg gccacaagtt cagcgtgcgc ggcgagggcg
      721 agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      781 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
      841 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      901 aggagcgcac catcagcttc aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt
      961 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1021 gcaacatcct ggggcacaag ctggagtaca actttccgcc gcccaactct actgatccag
     1081 aaggtgcata cgaaaccgtg aagaaggagt acaaacgtaa atggaatatt gtatggctca
     1141 aaccactggg attcaacaat acgtatacgc ttggggttaa agacgaactg gcgaaacagt
     1201 ataaccttaa aaccttcagt gacttagcga aaatctcgga taagctgatt ctgggtgcaa
     1261 cgatgttctt tttagaaaat cccgatggtt acccaggcct gcaaaaactg tacaatttca
     1321 aattcaagca caccaaaagc atggacatgg gtattcgcta taccgccatt gataataacg
     1381 aagttcaggt aattgatgcc tgggccactg atggcttgct ggtgagccac aaattaaaaa
     1441 ttctggagga tgataaagcg ttcttcccgc cgtattatgc tgcccccatc atccgtcagg
     1501 atgtcttaga taagcatcct gaactgaagg acgtgctgaa caaactcgcg aatcaaattt
     1561 cactggaaga aatgcagaaa ctgaattaca aggtggacgg tgagggtcag gacccagcga
     1621 aagtagctaa ggagtttttg aaagagaaag gtttaattac gcgtgaacaa aaactcatct
     1681 cagaagagga tctgaatgct gtgggccagg acacgcagga ggtcatcgtg gtgccacact
     1741 ccttgccctt taaggtggtg gtgatctcag ccatcctggc cctggtggtg ctcaccatca
     1801 tctcccttat catcctcatc aaatacttgc agaagaggcg ggaaaggcga cgccaggaga
     1861 acgctaactc cttttgctat gaaaatgagg tagccctgta ggcggcctat cgataatcaa
     1921 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1981 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     2041 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     2101 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2161 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     2221 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2281 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     2341 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2401 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2461 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     2521 cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     2581 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     2641 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     2701 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     2761 ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt
     2821 aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2881 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2941 ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3001 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3061 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3361 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3601 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3841 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3901 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3961 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4021 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4081 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4141 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4201 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4261 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4321 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4381 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4441 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4501 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4561 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4621 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4681 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4741 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4801 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4861 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4921 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     4981 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     5041 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     5101 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     5161 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     5221 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     5281 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     5341 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     5401 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5461 cttcgctatt acgccagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
     5521 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     5581 tggccaactc catcactagg ggttccttgt agttaatgat taacccgcca tgctacttat
     5641 ctacgtagcc atgctctagg aagatctctg cagagggccc tgcgtatgag tgcaagtggg
     5701 ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc ccgacccact
     5761 ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg
     5821 aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc
     5881 gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact
     5941 cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg
     6001 cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg
     6061 ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc
     6121 gtgcctgaga gcgcag
//
